Genes of virulence and antimicrobial resistance in Vibrio parahaemolyticus prevalent in areas of ostreiculture
DOI:
https://doi.org/10.20950/1678-2305.2018.364Keywords:
estuaries, antimicrobials, anthropogenic action, oystersAbstract
This study aimed to quantify Vibrio spp. and evaluate the profile of antimicrobial susceptibility and virulence factors in Vibrio parahaemolyticus strains in the samples of oysters collected from two estuaries of the Baixo Sul, Bahia. The samples were collected between June 2015 and January 2016 from natural banks (E1) (oyster) and from cultivation areas (E2) (water and oyster). For the quantification of Vibrio spp., the most probable number (MPN) was determined and the characterization of V. parahaemolyticus marker (species-specific) gene tl was performed. Pathogenicity was observed with the Kanagawa test, and the presence of the tdh, trh and ure genes were tested. The antimicrobial sensitivity test included disc diffusion method with 15 antimicrobial discs, β-lactamase enzyme production, and the presence of blaTEM, blaSHV, and blaCTX-M. The maximum density of Vibrio spp. in the samples from cultivation was 4.70 log MPN g-1 and extractivism was 6.10 log MPN g-1, with the temperature being the most influencing variable on the presence of microorganisms in the cultivation area (E2). The tl gene was detected in 71% of the isolates, without the presence of tdh, trh and ure genes. All strains of V. parahaemolyticus were Kanagawa negative. High antimicrobial resistance was observed in the β-lactam antibiotics (cephalothin - 72%, ampicillin - 60%) and aminoglycosides (amikacin - 64%), with multi-resistance in 88% of the strains of V. parahaemolyticus, of which 68% of the resistance was mediated by plasmids. Phenotypically, no production of β-lactamase enzymes was observed, but the presence of blaTEM genes was observed. The multiresistant character of plasmids reported in V. parahaemolyticus strains increases the concern about native bacteria in the marine environment since it can potentially compromise the control of this bacterium infection in bivalve mollusks.
References
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BEJ, A.K.; PATTERSON, D.P.; BRASHER, C.W.; VICKERY, M.C.L.; JONES, D.D.; KAYSNER, C.A. 1999 Detection of total and hemolysin producing Vibrio parahaemolyticus in shellfish using multiplex PCR amplification of tl, tdh and trh. Journal of Microbiological Methods, 36(3): 215-225.
BONDIOLI, A.C.V.; MARQUES, R.C.; TOLEDO, L.F.A.; BARBIERI, E. 2107 PCR-RFLP for identification of the pearl oyster Pinctada imbricate from Brazil and Venezuela. Boletim do Instituto de Pesca, 43(3): 459-463.
BOUKI, C.; VENIERI, D.; DIAMADOPOULOS, E. 2013 Detection and fate of antibiotic resistant bacteria in wastewater treatment plants: A review. Ecotoxicology and Environmental Safety, 91(1): 1-9.
BRASIL, 1997 Ministério da Saúde. Secretaria de Vigilí¢ncia Sanitária. Portaria nº 451, de 19 de setembro de 1997. Regulamento técnico princípios gerais para o estabelecimento de critérios e padrões microbiológicos para alimentos. Diário Oficial da União, Brasília, 02 de julho de 1998. Seção 1.
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BRASIL, 2012 Ministério da Pesca e Aquicultura. Instrução Normativa Interministerial no 7, de 8 de maio de 2012. Diário Oficial da União, Brasília, 9 de maio de 2012. Seção 1.
BRASIL, 2018 Ministério da Saúde. Secretária de Vigilí¢ncia em Saúde. Análise Epidemiológica dos Surtos de Doenças Transmitidas por Alimentos no Brasil. Disponível em: <http://portalarquivos2.saude.gov.br/images/pdf/2018/janeiro/17/Apresentacao-Surtos-DTA-2018.pdf >. Acesso em: 04 abr. 2018.
BUSH, K.; JACOBY, G.A. 2010 Updated functional classification of b-lactamases. Minireview. Antimicrobial Agents and Chemotherapy, 54(3): 969-976.
CABURLOTTO, G.; GENNARI, M.; VALENTINA, G.H.I.D.I.N.I.; MARIA, C.T.; MARIA, M.L.L.E.O. 2009 Presence of T3SS2 and other virulence-related genes in tdh-negative Vibrio parahaemolyticus environmental strains isolated from marine samples in the area of the Venetian Lagoon, Italy. FEMS Microbiology Ecology, 70(3): 506-514.
CECCARELLI, D.; HASAN, N.A.; HUQ, A.; COLWELL, R.R. 2013 Distribution and dynamics of epidemic and pandemic Vibrio parahaemolyticus virulence factors. Frontiers in Cellular in Infection Microbiology, 3(97): 1-9.
CDC - CENTERS FOR DISEASE CONTROL. 1998 Outbreak of Vibrio parahaemolyticus infections associated with eating raw oysters - pacific northwest, 1997. Morbidity and Mortality Weekly Report, 47(22): 457-462.
CLSI - CLINICAL AND LABORATORY STANDARDS INSTITUTE. 2010 Performance standards for antimicrobial susceptibility testing: seventeenth informational supplement. Wayne: Supplement M100-S20.
CLSI - CLINICAL AND LABORATORY STANDARDS INSTITUTE. 2009 Performance Standards for Antimicrobial Susceptibility Testing; Table: M100 - S19; 940 West Valley Road, Suite 1400 Wayne, PA 19087-1898 USA, 2009.
COSTA, R.A.; VIEIRA, G.H.; VIEIRA, R.H.S.; SAMPAIO, S.S. 2009 Vibrio em amostras de água de viveiros de cultivo do camarão marinho Litopenaeus vannamei, no Ceará-Brasil. Atlí¢ntica, 31(2): 177-182.
COSTA SOBRINHO, P.S.; DESTRO, M.T.; FRANCO, B.D.G.M.; LANDGRAF, M. 2011 Occurrence and distribution of Vibrio parahaemolyticus in retail oysters in Sao Paulo State, Brazil. Food Microbiology, 28(1): 137-140.
ELLINGTON, M.J.; KISTLER, J.; LIVERMORE, D.M.; WOODFORD, N. 2007 Multiplex PCR for rapid detection of genes encoding acquired metallo-β-lactamases. Journal of Antimicrobial Chemotherapy, 59(2): 321-2.
FISCHER, F.; NASCIMENTO, A.; PIERINE, C.; FISCHER, C.M.; FISCHER, F.; ROCHA, L.; MATOS, L.B.; SANTANA. L.; VINHARES, L.; SANTOS, M.E.P.; BRITO, M.R.; SANTOS FILHO, N.G. 2007 Baixo Sul da Bahia: uma proposta de desenvolvimento territorial. Projeto Série Editorial CIAGS, Salvador.
GUTIERREZ-WEST, C.K.; KLEIN, S.L.; LOVELL, C.R. 2013 High Frequency of virulence factor genes tdh, trh, and tlh in Vibrio parahaemolyticus strains isolated from a Pristine Estuary. Applied and Environmental Microbiology, 79(7): 2247-2252.
IBAM - INSTITUTO BRASILEIRO DE ADMINISTRAÇÃO MUNICIPAL. 2010 Estudo de caso: organização socioprodutiva de lideranças jovens em áreas rurais e estuarinas do território do Baixo Sul da Bahia. p. 53, Foro Iberoamericano y del Caribe - Melhores Práticas. Disponível em: <http://www.ibam.org.br/media/arquivos/estudos/estudo_terragua.pdf> Acesso em: 30 jul. 2015.
IBGE - INSTITUTO BRASILEIRO DE GEOGRAFIA E ESTATÍSTICA. 2015 Infográficos: Dados gerais do município de Taperoá. Disponível em: <http://http://cidades.ibge.gov.br/painel/painel.php?codmun=293120> Acesso em: 30 jul. 2015.
KANG, C.H.; SHIN, Y.; JANG, S.; KIM, S.; AN, S.; PARK, K.; SO, JS. 2017 Characterization of Vibrio parahaemolyticus isolated from oysters in Korea: Resistance to various antibiotics and prevalence of virulence genes. Marine Pollution Bulletin, 118(1-2):261-266.
KLEIN, S.L.; WEST, C.K.G.; MEJIA, D.M.; LOVELL, C.R. 2014 Genes similar to the Vibrio parahaemolyticus virulence-related genes tdh, tlh, and vscC2 occur in other Vibrionaceae species isolated from a pristine estuary. Applied and Environmental Microbiology, 80(2): 595-602.
KRUMPERMAN, P.H. 1983 Multiple antibiotic indexing of Escherichia coli to identify high risk sources of fecal contamination of foods. Applied and Environmental Microbiology, 46(1): 165-170.
LEAL, N.C.; SILVA, S.C.; CAVALCANTI, V.O.; FIGUEIROA, A.C.T.C.; NUNES, V.V.F.; MIRALLES, I.S.; HOFER, E. 2008 Vibrio parahaemolyticus serovar O3:K6 gastroenteritis in northeast Brazil. Journal of Applied Microbiology, 105(3): 691-697.
LEE, J. K.; JUNG, D. W.; EOM, S. Y.; OH, S. W.; KIM, Y.; KWAK, H. S.; KIM, Y. H. 2008 Occurrence of Vibrio parahaemolyticus in oysters from Korean retail outlets. Food Control, 19(10): 990-994.
LOU, Y.; HAIQUAN, L.; ZHANG, Z.; PAN, Y.; ZHAO, Y. 2016 Mismatch between antimicrobial resistance phenotype and genotype of pathogenic Vibrio parahaemolyticus isolated from seafood. Food Control, 59(1): 207-211.
MANJUSHA, S.; SARITA, G.B. 2013 Characterization of plasmids from multiple antibiotic resistant Vibrios isolated from molluscan and crustacean of Kerala. International Food Research Journal, 20(1): 77-86.
MEYER, G.; PICOLI, S.U. 2011 Fenótipos de betalactamases em Klebsiella pneumoniae de hospital de emergeÌ"šncia de Porto Alegre. Journal Brasileiro de Patologia e Medicina Laboratorial, 47(1): 25-31.
MOLINA-AJA, A.; GARCÍA-GASCA, A.; ABREU-GROBOIS, A.; BOLÁN-MEJÍA, C.; ROQUE, A.; GOMEZ-GIL, B. 2002 Plasmid profiling and antibiotic resistance of Vibrio strains isolated from cultured penaeid shrimp. FEMS Microbiology Letters, 213(1): 7-12.
NASCIMENTO, V.A.; MITTARAQUIS, A.S.P.; TRAVÁLIA, B.M.; SANTOS, R.C.A.; NUNES, M.L.; AQUINO, L.C.L. 2011 Qualidade Microbiológica de moluscos bivalves - sururu e ostras submetidos a tratamento térmico e estocagem congelada. Scientia Plena, 7(4): 1-5.
NOGUEROLA, I.; BLANCH, A.R. 2008 Identification of Vibrio spp. with a set of dichotomous keys. Journal of Applied Microbiology, 105(1): 175-185.
ORBAN, E.; DI LENA, G.; NEVIGATO, T.; CASINI, I.; CAPRONI, R.; SANTARONI, G.; GIULINI, G. 2006 Nutricional and commercial quality of the striped venus clam, Chamelea gallina, from the Adriatic sea. Food Chemistry, 101(3): 1063-1070.
ROCHA, R.S.; SOUSA, O.V.; VIEIRA, R.H.S.F. 2016 Multidrug-resistant Vibrio associated with an estuary affected by shrimp farming in Northeastern Brazil. Marine Pollution Bulletin, 105(1): 337-340.
ROJAS, M.V.R.; MATTÉ, M.H.; DROPA, M.; SILVA, M.L.; MATTÉ, G.R. 2011 Characterization of Vibrio parahaemolyticus isolated from oysters and mussels in São Paulo, Brazil. Revista do Instituto de Medicina Tropical de São Paulo, 53(4): 201-5.
SANDE, D.; MELO, T.A.; OLIVEIRA, S.A.; BARRETO, L.; TALBOT, T.; BOEHS, G.; ANDRIOLI, J.L. 2010 Prospecção de moluscos bivalves no estudo da poluição dos rios Cachoeira e Santana em Ilhéus, Bahia, Brasil. Brazilian Journal of Veterinary Research and Animal Science, 47(3): 190-196.
SANTOS, S.S.; BARRETO, L.M.; SILVEIRA, C.S.; REIS, N.A.; LIMA, K.A.; SOUZA, J.S.S.; EVANGELISTA-BARRETO, N.S. 2015 Condições sanitárias de ostras produzidas e comercializadas em Taperoá, Bahia e o efeito da depuração na redução da carga microbiana. Actapesca, 3(2): 49-60.
SHAW, K.S.; GOLDSTEIN, R.E.R.; HE, X.; JACOBS, J.M.; CRUMP, B.C.; SAPKOTA, A.R. 2014 Antimicrobial susceptibility of Vibrio vulnificus and Vibrio parahaemolyticus recovered from recreational and commercial areas of Chesapeake Bay and Maryland Coastal Bays. PloS One, 9(2): e89616.
SILVA, N.; JUNQUEIRA, V.C.A.; SILVEIRA, N.F.A.; TANIWAKI, M.H.; SANTOS, R.F.S.; GOMES, R.A.R. 2010 Manual de métodos de análise microbiológica de alimentos. São Paulo: Varela. 632p.
SILVA, C.C.; SILVA, J.C. 2007 Cultivo de ostras í Dossiê técnico. REDETEC Rede de Tecnologia do Rio de Janeiro. Disponível em:
< http://www.respostatecnica.org.br/dossie-tecnico/downloadsDT/MzA4>. Acesso em: 15 mar. 2018.
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2018-12-25
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